Scribd
Upload
73 views15 pages

Protein Folding

The document discusses biopolymers and protein folding and stability. It notes that heteropolymers form distinct folded structures through template-driven synthesis, while homopolymers form random collapsed structures. It describes the thermodynamics hypothesis that native protein structures reside in a global free energy minimum. Protein folding kinetics may result in trapped kinetic products differing from the native structure. It also discusses using denaturants like heat, urea or pH changes to perturb the equilibrium between folded and unfolded states to study protein folding transitions.

Uploaded by

bing_boom
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
73 views15 pages

Protein Folding

The document discusses biopolymers and protein folding and stability. It notes that heteropolymers form distinct folded structures through template-driven synthesis, while homopolymers form random collapsed structures. It describes the thermodynamics hypothesis that native protein structures reside in a global free energy minimum. Protein folding kinetics may result in trapped kinetic products differing from the native structure. It also discusses using denaturants like heat, urea or pH changes to perturb the equilibrium between folded and unfolded states to study protein folding transitions.

Uploaded by

bing_boom
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
You are on page 1/ 15

Biopolymers, Synthetic Polymers

Heteropolymers Form distinct folded, structures Template-driven synthesis , identical product

Homopolymers Random collapsed structures

Protein Folding & Stability


Helps you handle proteins appropriately Mutants are often folded differently than native forms Understanding molecular basis of diseases

Protein Folding & Stability


Protein folding can occur in the absence of additional manipulations (heating, pH)
Structures specified by amino acid sequence

Protein Folding & Stability


Thermodynamics hypothesis for protein folding
Native protein structures reside at a global free energy minimum Protein stability determined by G (folded, unfolded)

Protein Folding & Stability


Protein folding can occur in the absence of additional manipulations (heating, pH)
Structures specified by amino acid sequence

Thermodynamics hypothesis for protein folding


Native protein structures reside at a global free energy minimum

Protein folding kinetics


Trapped kinetic products that differ from natively folded structure

Monitoring protein folding, unfolding transitions


Equilibrium, reversible Two methods
Unfolded protein Observer protein folding Folded protein Denature Unfolded Observe protein folding

Denaturant, heat, pH Biophysical probes of protein structure


Circular dichroism

Circular dichroism
Folded (aqueous buffer)
270-290nm Unfolded (urea) tyrosine

tryptophan 280-300nm

Near UV CD spectrum

Chromopores: aromatic amino acids, disulphide bonds Due to absorption, dipole orientation, nature of surrounding environment

phenylalanine 250-270nm

Second law of thermodynamics and Entropy


An isolated system will always approach state of higher entropy (i.e. more disorganized) Unfolded protein chain has higher entropy than folded protein Protein folding in water
Higher entropy in water Water in contact with hydrophobic region, entropy of water decrease Hide hydrophobic regions to increase entropy of water

Folded Hydrophobic collapse (MAJOR FORCE) Intramolecular H-bonding (water)

Unfolding Configurational entropy (01.2kcal/mol/residue) H-bonding to solvent

Van der Waals interaction


Unfolded polypeptides exposes nonpolar groups Results in formation of clathrates (ordered solvent cages) NOT entropically favorable Hydrophobic effect increases solvent entropy due to release of (water) molecules (entropy of solvent INCREASES)

Unfolded Protein
Negative G Folding is spontaneous, unfolding occurs slowly Perturb equilibrium to favor unfolded state G thermodynamically
Lower Gunfolded Increase Gfolded Urea, heat, pH, guanidinium HCl native conditions

U U
rxn coord

denaturing conditions

FOLDED active

8M urea (denaturant) 2ME (reducing S-S)

UNFOLDED inactive

The free energy change on folding or unfolding is due to the combined effects of both protein folding/unfolding and hydration changes. These compensate to such a large extent that the free energy of stability of a typical protein is only 40-90 kJ mol-1 (equivalent to very few hydrogen bonds), whereas the enthalpy change (and temperature times the entropy change) may be greater than 500 kJ mol-1 different.

Perturb equilibrium
Increase temperature

Two-state model of folding

fraction unfolded

U
[U] = [F]

0.5

F
[GdmHCl]

Thermodynamic hypothesis
Protein fold into their lowest energy state

Kinetic trap
Energy barrier to lowest energy state (disulphide bonds, low pH)

Folding pathways
Random or directed

Chemical denaturants to lower free energy of unfolded state to populate enough to observe transition

You might also like